pSP-luc+NF Fusion Vector Technical Bulletin

Literature # TB210

The pSP-luc+NF Fusion Vector is a luciferase cassette vector containing the engineered firefly luciferase gene, luc+NF. The luc+NF gene is related to the luc+ gene found in our pGL3 family of eukaryotic reporter vectors but has been further modified for maximum flexibility in constructing N-terminal fusions (NF) with luciferase. The pSP-luc+NF Fusion Vector is not intended for expression of luciferase in eukaryotic cells because it does not contain eukaryotic promoters, enhancers or polyadenylation signals.

The luc+NF gene is positioned downstream of an SP6 promoter and a ribosome binding site. An opposing T7 promoter is located immediately downstream of luc+NF. Thus, the pSP-luc+NF Fusion Vector provides a convenient template for in vitro synthesis of both sense and anti-sense luciferase transcripts for studies involving in situ hybridization, RNA processing, RNA transfection or coupled in vitro transcription/translation and protein folding. Multiple cloning regions containing recognition sequences for commonly used restriction enzymes are positioned at the 5´ and 3´ ends of luc+NF to provide maximum flexibility in cloning. Subcloning luc+NF into expression vectors provides a useful genetic reporter with exceptional sensitivity. Luciferase enzymatic activity can be assayed most efficiently using the Luciferase Assay System.